Dendritic cells (DCs) are unique antigen presenting cells that are used in cancer immunotherapy, with more than 20 years of clinical trials in this field. The production of clinical grade monocyte-derived DCs is highly desired and globally performed, with a large space for improvement and development of clinical standard operating procedures (CSOP). This process is highly dependent on three constraints: used cytokines for cell differentiation, maturating agents and culture medium. There is a clear lack of studies focusing on how the used culture medium and its composition and components affect the process of ex-vivo generation of monocyte-derived DCs. This study aims to compare the effect of 4 culture media (3 GMP media and 1 medium widely used in research) during monocyte-derived DCs differentiation.
We characterized DC viability, differentiation, maturation, internalization of tumor lysates, cytokines production and autologous T cell stimulatory capacity, as well as metabolomic profiles. Commercially available culture media for clinical use were tested, namely DendriMACS, AIM-V and X-VIVO 15. RPMI was also tested as a comparative term given that it is largely used in pre-clinical research.
In terms of differentiation, maturation, DC uptake capacity, and metabolic profiles, AIM-V and X-VIVO 15 present similar results. However, the use of X-VIVO 15 shows an enhanced DC production of IL-12. DCs cultured in X-VIVO 15 and AIM-V media are able to induce a superior stimulation of T cells, mainly CTLs and Th1 responses, while DCs cultured in DendriMACS are more prone to induce Treg polarization.
Our data show that X-VIVO 15 and AIM-V culture media are preferable to support the differentiation of DC to be used in immunostimulatory approaches such as in cancer cell therapy. Overall, this study highlights the need of previously and carefully defining the culture medium to be used in DC cancer immunotherapy, for a better aim to the therapeutic goal and potentiating the clinical experiment and final patient output.
Grupo Tecnimede.
This study received funding from the project ImmunoDCs@CancerStemCells: Cellular Immunotherapy towards the elimination of cancer stem cells (Ref.: POCI-01-0247-FEDER-033532), co-funded by the European Regional Development Fund (FEDER), Competitiveness and Internationalization Operational Program (COMPETE2020) and Own Revenues of the University of Coimbra. João Calmeiro is supported by the Portuguese Science and Technology Foundation (FCT) through an individual PhD fellowship (PD/BDE/135076/2017).
All authors have declared no conflicts of interest.